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Application of indirubin for preparing medicine for preventing blood vessel from regenerating

An angiogenesis and drug technology, applied in the application field of indirubin in the preparation of angiogenesis-inhibiting drugs, can solve the problems of no indirubin inhibiting angiogenesis drugs, no indirubin inhibiting angiogenesis and the like

Inactive Publication Date: 2008-10-15
EAST CHINA NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although it has been reported that indirubin can directly act on tumor cells to have an anticancer effect, there is no report that indirubin inhibits angiogenesis, or inhibits tumors by inhibiting angiogenesis, and there is no indirubin as an inhibitor of angiogenesis. new drug reports

Method used

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  • Application of indirubin for preparing medicine for preventing blood vessel from regenerating
  • Application of indirubin for preparing medicine for preventing blood vessel from regenerating
  • Application of indirubin for preparing medicine for preventing blood vessel from regenerating

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Indirubin inhibits the proliferation of human umbilical vein endothelial cells and human microvascular endothelial cells

[0023] Purpose and principle: Endothelial cell proliferation experiment adopts MTS assay. The MTS assay is a colorimetric method that indirectly measures the number of living cells. In living cells, the NADPH dehydrogenase in the mitochondria converts the MTS tetrazolium salt compound into a colored substance that is dissolved in the cell culture fluid. The more the number of living cells, the more the tetrazolium salt can be converted into this colored substance. Therefore, measuring the light absorption value of the substance at 490nm can indirectly reflect the number of living cells. This method can be used to evaluate the effects of drugs on the proliferation of two endothelial cells.

[0024] Method: Insert endothelial cells (human umbilical vein endothelial cells (HUVEC) or human microvascular endothelial cells (HMEC-1) into a 96-well p...

Embodiment 2

[0026] Example 2: Indirubin inhibits the migration of human umbilical vein endothelial cells from scratches in vitro

[0027] Purpose and principle: To evaluate the effect of drugs on the migration ability of endothelial cells by observing, recording, and counting the number of newly migrated endothelial cells in the scratch with or without the action of drugs.

[0028] Method: Insert human umbilical vein endothelial (HUVEC) cells into a 6-well plate. When the cells grow to logarithmic phase, cross the lines with the tip of a micropipette, wash the marked cells with PBS, and add different drug concentrations And 4ng / mlVEGF medium, in 5% CO 2 The cells cultured at 37°C in the incubator on both sides of the streak migrate to the center of the streak area.

[0029] Results and evaluation: Observing and taking statistics under the OLYMPUS inverted microscope, the results are: A is the control group; B is the indirubin 100nM group. Compared with the control group, after adding indirubi...

Embodiment 3

[0030] Example 3: Indirubin inhibits human umbilical vein endothelial cell modification Boyden cell migration experiment

[0031] Purpose and principle: Boyben chamber was originally invented by Boyden to study the chemotaxis of white blood cells. This research is based on the chamber being separated into two parts by a micropore. Cells are placed on the upper part and the lower part contains chemokines. Under the induction of chemokines, the upper layer can migrate through the microporous membrane to the bottom of the membrane and attach to the back of the membrane. The Boyden chamber migration experiment of human umbilical vein endothelial cells used Millicell (Millipore) cell culture chamber. Under the induction of growth factors, cells can pass through a polycarbonate membrane with a pore size of 8μm. This experiment can detect the effect of drugs on the ability of endothelial cells to pass through the microporous membrane.

[0032] Method: Coat the cell with 0.1% gelatin, 5% ...

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Abstract

The invention relates to a new application of indirubin, particularly to an application of the indirubin in preparing drugs with angiogenesis inhibiting effect for treating angiogenesis diseases such as eye diseases, tumors, arthritis, hemangioma and psoriasis. In human microvascular endothelial cell-1 (HMEC-1) proliferation test, human umbilical vein endothelial cell (HUVEC) proliferation test, HUVEC in vitro scratch migration test, HUVEC-modified Boyden chamber migration test, HMEC-1 canalization test and chick embryo allantoic membrane test, the indirubin has evident inhibitory effect. Therefore, the indirubin can be used for preparing drugs with angiogenesis inhibiting effect.

Description

Technical field [0001] The invention relates to a new use of indirubin, in particular to the application of indirubin in preparing drugs for inhibiting angiogenesis. Background technique [0002] Angiogenesis is the process of generating new blood vessels on the basis of existing blood vessels, including five basic processes: ①Degradation of vascular basement membrane; ②Endothelial cells migrate through the basement membrane to the surrounding matrix of blood vessels; ③Endothelial cells proliferate, adhere to and connect to each other; ④Form a tube-like structure; ⑤Matrix remodeling, smooth muscle cell wrapping and blood vessel anastomosis form a vascular network. [0003] Angiogenesis occurs under both physiological and pathological conditions. Physiological angiogenesis mainly occurs in embryonic development and female physiological cycle, which is very important for normal development. Pathological angiogenesis is an important cause of many major diseases, including tumors, Ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/404A61P35/00A61P29/00A61P27/02A61P43/00
Inventor 易正芳张晓丽赖力张京卢彬彬易庭芳刘明耀
Owner EAST CHINA NORMAL UNIV
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